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In recent years, the presence of molecules derived from aromatic amino acids in wines has been increasingly demonstrated to have a significant influence on wine quality and stability. In addition, interactions between different yeast species have been observed to influence these final properties. In this study, a screening of 81 yeast strains from different environments was carried out to establish a consortium that would promote the improvement of indolic compound levels in wine. Two strains, Saccharomyces uvarum and Saccharomyces eubayanus, with robust fermentative capacity were selected to be combined with a Saccharomyces cerevisiae strain with a predisposition towards the production of indolic compounds. Fermentation dynamics were studied in pure cultures, co-inoculations and sequential inoculations, analysing strain interactions and end-of-fermentation characteristics. Fermentations showing significant interactions were further analyzed for the resulting indolic compounds and aroma profile, with the aim of observing potential interactions and synergies resulting from the combination of different strains in the final wine. Sequential inoculation of S. cerevisiae after S. uvarum or S. eubayanus was observed to increase indolic compound levels, particularly serotonin and 3-indoleacetic acid. This study is the first to demonstrate how the formation of microbial consortia can serve as a useful strategy to enhance compounds with interesting properties in wine, paving the way for future studies and combinations

Hydroxytyrosol (HT) is a well-known compound for its bioactive properties. It is naturally present in olives, olive oil, and wine. Its presence in wines is partly due to its production during alcoholic fermentation by yeast through a hydroxylation of tyrosol formed through the Ehrlich pathway. This work aims to explore the influence of yeast assimilable nitrogen (YAN) and glucose content as precursors of HT formation during alcoholic fermentation. Commercial Saccharomyces cerevisiae QA23 and its metabolically engineered strain were used to ferment synthetic must. Each strain was tested at two different YAN concentrations (210 and 300 mg L−1) and two glucose concentrations (100 and 240 g L−1). This work confirms that the less YAN and the more glucose, the higher the HT content, with fermentations carried out with the metabolically engineered strain being the ones with the highest HT content (0.6 mg L−1).

The dataset include the raw data of the analysys of HPLC-MS/MS for tyrosol and hydroxytyrosol. The samples were from fermented synthetic must by the yeas strain Red Fruit in different conditions of YAN (210 mg/L and 300 mg/L) and different glucose conditions (100 g/L y 240 g/L).

Yeasts are feasible and efective bioreactors and, therefore, there is a great interest in their industrial employment for the production of a wide range of molecules. In this study, the production by Saccharomyces cerevisiae of bioactive compounds such as hydroxytyrosol (HT), tyrosol (TYR) and melatonin (MEL) vs. volatile compounds in fermented must was studied. The concentration of the bioactive compounds HT and MEL in fermented must employing diferent yeast strains revealed that the higher the concentrations, the lower the amount of volatile compounds determined. This inverse correlation was especially remarkable with respect to the production of higher alcohols, especially 2-phenylethanol (2-PE) and esters. Furthermore, the employment of a modifed Aro4pᴷ²²⁹ᴸ S. cerevisiae QA23 yeast strain which overproduces HT, gave rise to fermented must also higher in 2-PE and their corresponding esters but with an outstanding less presence of other important esters such as ethyl hexanoate and ethyl octanoate. Both premises could point out that S. cerevisiae might have diferent approaches to handling cell stress/toxicity due to their nitrogen metabolism. One detoxifying pathway could be through the production of higher alcohols and these in turn to esters and the other be more related to synthesizing antioxidant molecules such as MEL and HT.

Las bebidas fermentadas son una fuente de compuestos bioactivos, entre ellos derivados del metabolismo de aminoácidos aromáticos. Sin embargo, el conocimiento de las rutas que intervienen en la síntesis de estas moléculas bioactivas por parte de las levaduras durante la fermentación es limitado. Mejorar el conocimiento de las rutas implicadas puede proporcionar información para incrementar el contenido en compuestos bioactivos en bebidas fermentadas. Además, el estudio de los mecanismos de acción por los cuales estos compuestos bioactivos ejercen su efecto biológico resulta de interés para poder profundizar en el conocimiento de sus propiedades saludables. En la presente Tesis Doctoral se ha realizado un estudio de las rutas metabólicas relacionadas con la síntesis de melatonina (MEL) y hidroxitirosol (HT) en la levadura mediante el uso de sus precursores aminoácidos aromáticos (triptófano (L-TRP) y tirosina (TYR), respectivamente) marcados isotópicamente. En el análisis por UHPLC/HRMS se detectaron serotonina (SER), N-acetilserotonina (NAS) y MEL marcados. No obstante, la presencia de los compuestos mencionados no marcados muestra que durante la fermentación alcohólica la síntesis de MEL por parte de la levadura ocurre tanto a partir del L-TRP del mosto como del L-TRP sintetizado por la levadura. De igual forma, en fermentaciones con el aminoácido TYR isotópicamente marcado, se detectaron ácido 4-hidroxifenilacético, tirosol (TYRS) y HT marcados y sin marcar. Por consiguiente, la ruta de Ehrlich no es la mayoritaria para la producción de HT por parte de la levadura durante la fermentación alcohólica. Asimismo, se ha llevado a cabo la evaluación de la ingesta dietética del HT en la población adulta de la Unión Europea (UE) y la contribución de sus principales fuentes a su ingesta. Los estudios realizados sobre la ingesta dietética del HT muestran que Grecia, España, Dinamarca, Chipre e Italia son los países que presentan la mayor ingesta de HT siendo la aceituna de

Introducción: según el Instituto Nacional de Estadística un 36,4% de personas mayores de 15 años se declaró sedentaria. Una de las razones de esta inactividad física se podría atribuir al nuevo modo de vida, envuelto en tecnología y cada vez actividades más estáticas. Sin embargo, la tecnología como el móvil puede ser una herramienta fundamental para cambiar esto, por su accesibilidad y uso por parte de la población. Objetivo: promover los hábitos saludables mediante herramientas tecnológicas. En concreto, crear un cuestionario de preguntas test en relación con la nutrición y la actividad física que permita a los usuarios aumentar su conocimiento. Además, se pretende, evaluar dichas preguntas, mediante una encuesta de usabilidad que será facilitada a los participantes y valorar la eficacia de la gamificación. Materiales y método: realización de una revisión bibliográfica de documentos relacionados con la gamificación y actividad física de la base de datos Scopus. Creación, uso y distribución de un elemento de gamificación, en nuestro caso un cuestionario, dirigido a la población adulta. Evaluación de la población y resultados mediante encuestas. Resultados: participaron 29 personas de entre 18-76 años. La puntuación media de acierto de los test fue de 57, 63 y 63, siendo el tiempo medio cada vez menor. La encuesta de usabilidad demostró en la mayoría de las preguntas una media de 7-8 sobre 9. Conclusión: la revisión bibliográfica reveló resultados positivos hacia la gamificación, al igual que nuestros resultados, los cuales, demostraban una visión favorable hacia nuestro elemento de gamificación por parte de la población de estudio.

Yeasts can synthetise bioactive compounds such as Melatonin (MEL), Serotonin (SER) and Hydroxytyrosol (HT). Deciphering the mechanisms involved in their formation can lead to exploit this fact to increase the bioactive potential of fermented beverages. Quantitative analysis using labelled compounds, 15-N2 l-tryptophan and 13-C tyrosine, allowed tracking the formation of the above-mentioned bioactive compounds during the alcoholic fermentation of synthetic must by two different Saccharomyces cerevisiae strains. Labelled and unlabelled MEL, SER and HT were undoubtedly identified and quantified by High Resolution Mass Spectrometry (HRMS). Our results prove that there are at least two pathways involved in MEL biosynthesis by yeast. One starts with tryptophan as precursor being known for the vertebrates’ pathway. Additionally, MEL is produced from SER which in turn is consistent with the plants’ biosynthesis pathway. Concerning HT, it can be formed both from labelled tyrosine and from intermediates of the Erlich pathway.

Promoting a healthy diet is a relevant strategy for preventing non-communicable diseases. This study aims to evaluate the impact of an innovative tool, the SAlBi educa nutrition app, in primary healthcare dietary counseling to improve dietary profiles as well as adherence to the Mediterranean diet. A multi-center randomized control trial comprising 104 participants was performed. Both control (n = 49) and intervention (n = 55) groups attended four once-weekly sessions focusing on healthy eating habits and physical activity, over one month. As well as attending the meetings, the intervention group used the app, which provides self-monitoring and tailored dietary advice based on the Mediterranean diet model. In a second intervention (one arm trial), the potential of SAlBi educa was evaluated for three months during the COVID-19 pandemic. At 4 weeks, the intervention group had significantly increased their carbohydrate intake (7.7% (95% CI: 0.16 to 15.2)) and decreased their total fat intake (−5.7% (95% CI: −10.4 to −1.15)) compared to the control group. Significant differences were also found for carbohydrates (3.5% (95% CI: −1.0 to 5.8)), total fats (−5.9% (95% CI: −8.9 to −3.0)), fruits and vegetables (266.3 g/day (95% CI: 130.0 to 402.6)), legumes (7.7g/day (95% CI: 0.2 to 15.1)), starchy foods (36.4 g/day (95% CI: 1.1 to 71.7)), red meat (−17.5 g/day (95% CI: −34.0 to −1.1)), and processed meat (−6.6 g/day (95% CI: −13.1 to −0.1)) intakes during the COVID-19 pandemic. SAlBi educa is a useful tool to support nutrition counseling in primary healthcare, including in special situations such as the COVID-19 pandemic. Trial registration: ISRCTN57186362.

In recent years, the use of applications to improve dietary habits has increased. Although numerous nutrition apps are available on the market, only few have been developed by health and nutrition professionals based on scientific evidence and subsequently tested to prove their usability. The main objective of this study was to design, develop, and evaluate the usability of a tailored nutrition application to be used to promote healthy eating habits. In order to decide app design and content, three focus groups took place with fifteen professionals from primary healthcare, nutrition, and food science and computer science, as well as expert users. For the general and feedback message design, a reference model based on the scientific literature was developed. To address the multi-perspective approach of users' and external healthcare professionals' feedback, a one-day pilot testing with potential users and healthcare professionals was conducted with four focus groups. To evaluate the relevance and potential usability of the app a 1-month pilot test was conducted in a real-life environment. A total of 42 volunteers participated in the one-day pilot testing, and 39 potential users participated in the 1-month pilot test. The SAlBi educa app developed includes an online dietary record, a self-monitoring tool to evaluate dietary patterns, general and feedback messages, and examples of traditional Mediterranean recipes. The usability study showed that volunteers think that SAlBi educa is pleasant (59%) and easy to learn to use (94%). Over 84% of the volunteers declared that the nutritional messages were clear and useful. Volunteers stated that general and tailored recommendations, as well as self-monitoring, were SAlBi educa's most motivating and useful features. SAlBi educa is an innovative, user-friendly nutritional education tool with the potential to engage and help individuals to follow dietary habits based on the Mediterranean model.

Hydroxytyrosol (HT) is a phenolic compound with proven biological properties present in a limited number of foods such as table olives, virgin olive oil (VOO) and wines. The present work aims to evaluate the dietary intake of HT in the European (EU) population by compiling scattered literature data on its concentration in foods. The consumption of the involved foods was estimated based on the EFSA Comprehensive European Food Consumption Database. The updated average contents of HT are as follows: 629.1, 5.2 and 2.1 µg/g for olives, olive oil and wine, respectively. The HT estimated intake in the European Union (EU) adult population falls within 0.13–6.82 mg/day/person, with table olives and wine being the main contributors. The estimated mean dietary intake of HT in EU countries is 1.97 ± 2.62 mg/day. Greece showed the highest HT intake (6.82 mg/day), while Austria presented the lowest (0.13 mg/day). Moreover, HT is an authorized novel food ingredient in the EU that can be added to different foods. Since the estimated HT intake is substantially low, the use of HT as a food ingredient seems feasible. This opens new possibilities for revalorizing waste products from olive oil and olive production which are rich HT sources.

Neuroinflammation is a common feature shared by neurodegenerative disorders, such as Parkinson’sdisease(PD),andseemstoplayakeyroleintheirdevelopmentandprogression. Microglia cells, the principal orchestrators of neuroinflammation, can be polarized in different phenotypes, which means they are able to have anti-inflammatory, pro-inflammatory, or neurodegenerative effects. Increasing evidence supports that the traditional Mediterranean dietary pattern is related to the reduction of cognitive decline in neurodegenerative diseases. A considerable intake of plant foods, fish, and extra virgin olive oil (EVOO), as well as a moderate consumption of red wine, all characteristicoftheMediterraneandiet(MD),arebehindtheseeffects. Thesefoodsareespeciallyrich in polyphenols, being the most relevant in the MD hydroxytyrosol (HT) and their derivatives present in EVOO, which have demonstrated a wide array of biological activities. Here, we demonstrate that HT is able to reduce the inflammation induced by two different stimuli: lipopolysaccharide and α-synuclein. We also study the possible molecular mechanisms involved in the anti-inflammatory effectofHT,includingthestudyofnuclearfactorkappaB(NF-κB),mitogen-activatedproteinkinases (MAPKs), nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, and inflammasome. Our data support the use of HT to prevent the inflammation associated with PD and shed light into the relationship between MD and this neurological disorder.

Actualmente, el exceso de peso está considerado un problema de salud pública a nivel mundial. El sobrepeso es la antesala a la obesidad; esta acumulación anormal de grasa corporal está considerada un factor de riesgo, asociado a la morbimortalidad por enfermedades crónicas no transmisibles. Las causas de esta tendencia creciente son los malos hábitos alimenticios y un aumento del sedentarismo; por lo tanto, para hacer frente a este problema es necesario fomentar y promocionar una alimentación saludable, basada en el modelo de Dieta Mediterránea, y aumentar la práctica de actividad física. Tanto a nivel nacional como internacional se han propuesto políticas alimentarias e intervenciones en el sistema sanitario para poner fin a esta epidemia de sobrepeso y obesidad. Este trabajo pretende analizar la eficacia del diseño y aplicación de talleres de Consejo Dietético en Atención Primaria para la mejora de hábitos de vida saludable. Durante el comienzo del presente año se realizó una intervención en dos centros de salud de la provincia de Sevilla. Los voluntarios reclutados recibieron cuatro talleres semanales, desarrollados ex profeso en este trabajo, impartidos por los profesionales sanitarios, sobre alimentación saludable y actividad física. Para la evaluación de la intervención se entregaron distintos cuestionarios que analizaban las mejoras de los participantes. En aquellas personas que permanecieron hasta el final del estudio hubo mejorías en la adherencia a la Dieta Mediterránea, en la práctica de actividad física, en conocimientos nutricionales adquiridos y en las cifras de peso; aunque el poco tiempo que duró la intervención (cuatro semanas) no fue suficiente para apreciar cambios importantes en los patrones dietéticos. El Consejo Dietético en Atención Primaria es una herramienta útil para mejorar los conocimientos de la población sobre estilos de vida saludable, pero la impartición únicamente de talleres puede no ser suficiente para conseguir cambios en los patr

The leaves of Vitis vinifera L. have been used for a long time in traditional medicine for the treatment of many ailments. Grape polyphenols, indeed, have been demonstrated to be able to defend against oxidative stress, responsible for various disorders such as cancer, diabetes and neurodegenerative diseases. The effects of different extraction techniques, Soxhlet (SOX), Accelerated Solvent (ASE 40, ASE 50) and Ultrasound Assisted Extraction (UAE) were studied in this work to evaluate their impact on the chemical profile and bioactive potential of Vitis vinifera L. (cv. Aglianico) leaf extracts. The phytochemical profile was investigated by HPLC-DAD and 9 phenolic compounds were identified and quantified in the extract. Moreover, the antioxidant, anticholinesterase and antityrosinase activities were evaluated. In detail, the total polyphenol content and antioxidant activity (2,2-diphenyl-1-picrylhydrazyl, Oxygen Radical Absorbance Capacities and β-Carotene Bleaching assays) were evaluated and compared to assess the Relative Antioxidant Capacity Index (RACI). To test the inhibitory activity of extracts towards cholinesterases, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition assays were performed. SOX and ASE 50 have shown the highest value of RACI, 0.76 and 0.65, respectively. Regarding enzymatic inhibitory activity, ASE 50 (IC50 = 107.16 ± 8.12 μg/mL) and SOX (IC50 = 171.34 ± 12.12 μg/mL) extracts exhibited the highest AChE and BChE inhibitory activity, respectively, while UAE (IC50 = 293.2 ± 25.6 μg/mL, followed by SOX (IC50 = 302.5 ± 38.3 μg/mL) showed the highest tyrosinase inhibition value. Our results demonstrated for the first time that Aglianico leaves are important sources of phenols that could be used to prevent oxidative stress and be potentially helpful in diseases treatable with tyrosinase and cholinesterase inhibitors, like myasthenia gravis or Alzheimer’s.

Hydroxytyrosol is well known for its potent antioxidant activity and anticarcinogenic, antimicrobial, cardioprotective and neuroprotective properties. Main food sources are olive oil (formed from the hydrolysis of oleuropein) and wine. One possible explanation to its origin in wines is the synthesis from tyrosol, which in turn is produced from the Ehrlich pathway by yeasts. This work aims to explore the factors that could increase the content as the strain of yeast, the initial tyrosine concentrations as precursor and the effect of synthetic and sterilized natural grape musts. Alcoholic fermentations in synthetic must showed that hydroxytyrosol is produced by all the yeast strains under study. Commercial Saccharomyces cerevisiae yeasts were those which produced higher concentrations, being the Red Fruit strain the biggest producer (6.12 ng/mL). Once the strain was selected, alcoholic fermentations were performed in synthetic must, with different tyrosine concentrations. The amount of hydroxytyrosol did not increase in a proportional way as tyrosine does. On the other hand, higher concentrations of hydroxytyrosol were obtained in natural grape musts (10.46 ng/mL) than in synthetic must (4.03 ng/mL). This work confirms the capacity of winemaking yeasts to produce the bioactive hydroxytyrosol.

Melatonin and serotonin are bioactive compounds present in foods and beverages and related to neuroprotection and anti-angiogenesis, among other activities. They have been described in wines and the role of yeast in their formation is clear. Thus, this study evaluates the content of these bioactives and other related indolic compounds in beer. For this purpose, commercial beers were analyzed by a validated UHPLC-HRMS method and sample treatment optimized due to the low concentrations expected. Moreover, a wort was fermented with different commercial beer yeast (Abbaye, Diamond, SafAle, SafLager) in order to monitor the formation of these bioactives during the elaboration process. Results show that indolic compounds such as N-acetylserotonin and 3-indoleacetic acid are produced during the alcoholic fermentation of wort. Moreover, the occurrence of four indolic compounds (5-hydroxytryptophan, N-acetylserotonin, 3-indoleacetic acid, L-tryptophan ethyl ester) in commercial beers is reported for the first time.

El dasaset incluye los datos brutos de los experimentos western blot y ELISA realizados en el trabajo titulado "Anti-VEGF Effect of Bioactive Indolic Compounds and Hydroxytyrosol Metabolites". También se incluyen las medias y las desviaciones estándar.

Angiogenesis drives evolution and destabilisation of atherosclerotic plaques and the growth and expansion of tumour cells. Vascular endothelial growth factor (VEGF) is the main endogenous pro-angiogenic factor in humans. The aim was to provide insight into the anti-VEGF activity of bioactive compounds derived from aromatic amino acids (serotonin, melatonin, 3-indoleacetic acid, 5-hydroxytryptophol and hydroxytyrosol). Experiments involved endothelial cell migration (wound-healing assay), the molecular mechanisms (ELISA assay) and the downstream effects (phospholipase C gamma 1 (PLCγ1), protein kinase B (Akt) and endothelial nitric oxide synthase (eNOS) by Western blot) on human umbilical vein endothelial cells (HUVECs). The data suggest for the first time that hydroxytyrosol interacts with surface components of the endothelial cell membrane (, preventing VEGF from activating its receptor. Serotonin and 5-hydroxytryptophol significantly inhibited HUVEC migration (98% and 50%, respectively) following the same mechanism. Conversely to other bioactive compounds, the anti-angiogenic effect of melatonin, serotonin, 3-indoleacetic acid and 5-hydroxytryptophol is not mediated via PLCγ1. However, hydroxytyrosol inhibits PLCγ1 phosphorylation. Additionally, melatonin and serotonin maintained eNOS phosphorylation and hydroxytyrosol significantly activated eNOS-all via Akt. These data provide new evidence supporting the interest in melatonin, serotonin, 3-indoleacetic acid, 5-hydroxytryptophol and hydroxytyrosol for their further exploitation as anti-VEGF ingredients in food.

Melatonin (Mel), originally considered a neurohormone, has been detected in beverages and food-fermented products in which yeast metabolism is highly important. This indolamine is synthesized from serotonin, with L-tryptophan being the initial substrate of both. Regarding Mel metabolism, the biosynthetic pathway in mammals consists in four-step reactions. However, six genes are implicated in the synthesis of Mel in plants, which suggest the presence of many pathways. The aim of this study was to provide new empirical data on the production of Mel and other indole-related compounds in the yeast Saccharomyces cerevisiae (S. cerevisiae). To this end, we performed the addition of the pathway intermediates in S. cerevisiae cells in different growth stages (exponential and arrested cells) to follow the bioconversion and new indolic compound production from them. The different bioconverted indolic compounds tested (L-tryptophan, 5-hydroxytryptophan, tryptamine, serotonin, N-acetylserotonin, 5-methoxytryptamine, and Mel) were analyzed by UHPLC-MS/MS from the extra- and intracellular contents. Our results showed that serotonin, in yeast, was prevalently formed via tryptophan decarboxylation, followed by tryptamine hydroxylation as in plants. Mel production from serotonin can be achieved by either N-acetylation, followed by O-methylation or O-methylation, in turn followed by N-acetylation. Accordingly, the classic pathway of Mel synthesis in vertebrates does not seems prevalent in yeast.

Certain bioactive compounds that derive from tryptophan have been determined in fermented foods, which suggests that the role of yeast is putative in origin. Melatonin is a neurohormone in humans that plays an important role in health. The activity of other compounds, such as 3-indoleacetic acid, has been recently highlighted, and interest in elucidating the conditions of their production has grown. However, the biosynthesis of melatonin by yeasts remains unclear to a large extent. Therefore, this work was undertaken to demonstrate the unequivocal synthesis of melatonin and other compounds that derive from tryptophan metabolism by yeast by determining them in the intracellular compartment. By high-resolution mass spectrometry and a validated method, tryptophan itself, melatonin, serotonin, N-acetyl-5-hydroxytryptamine and 3-indoleacetic acid, were identified in the intracellular compartment of Saccharomyces and non-Saccharomyces wine yeasts.

Background and Aims:Tryptophan is involved in the formation of bioactive compounds, such as melatonin (MEL) and3-indoleacetic acid (3-IAA), by yeast. Melatonin is a neurohormone whose occurrence in wine has been widely reported inrecent years. The occurrence, however, of MEL and other indolic compounds related to tryptophan metabolism by wineyeast strains has been scarcely reported in grape musts. This work examined the occurrence of these compounds during thealcoholic fermentation (AF) of musts from seven grape cultivars, Corredera, Chardonnay, Moscatel, Palomino Fino, Sau-vignon Blanc, Tempranillo and Vijiriega.Methods and Results:Must was fermented with threeSaccharomyces cerevisiaestrains and then in two cases an additionalsequential inoculation with the non-SaccharomycesyeastTorulaspora delbrueckiiwas carried out. Fermented must samples wereanalysed by UHPLC/HRMS to determine the concentration of:L-tryptophan, 5-hydroxytryptophan, 5-hydroxytryptamine,N-acetyl-5-hydroxytryptamine, MEL, 3-IAA, tryptamine, tryptophol andL-tryptophan ethyl ester. The profile of indoliccompounds during AF with the Aroma White strain depended on the cultivar. The yeast strain did not influence the profile ofindolic compounds; instead, fermentation time was found to be a more influential factor.Conclusions:The production of indolic compounds during the AF depends largely on the cultivar used and the day of fer-mentation on natural grape musts.Significance of the Study:This is thefirst study that quantifies 5-hydroxytryptophan andN-acetyl-5-hydroxytryptamineduring the AF of grape must. The occurrence of compounds with bioactive potential, for example 3-IAA and ML, during fer-mentation with commercial yeast strains is also described.

Neuroinflammation is a pathological feature of quite a number of Central Nervous System diseases such as Alzheimer and Parkinson’s disease among others. The hallmark of brain neuroinflammation is the activation of microglia, which are the immune resident cells in the brain and represents the first line of defense when injury or disease occur. Microglial activated cells can adopt different phenotypes to carry out its diverse functions. Thus, the shift into pro-inflammatory/neurotoxic or anti-inflammatory/neuroprotective phenotypes, depending of the brain environment, has totally changed the understanding of microglia in neurodegenerative disease. For this reason, novel therapeutic strategies which aim to modify the microglia polarization are being developed. Additionally, the understanding of how nutrition may influence the prevention and/or treatment of neurodegenerative diseases has grown greatly in recent years. The protective role of Mediterranean diet (MD) in preventing neurodegenerative diseases has been reported in a number of studies. The Mediterranean dietary pattern includes as distinctive features the moderate intake of red wine and extra virgin olive oil, both of them rich in polyphenolic compounds, such as resveratrol, oleuropein and hydroxytyrosol and their derivatives, which have demonstrated anti-inflammatory effects on microglia on in vitro studies. This review summarizes our understanding of the role of dietary phenolic compounds characteristic of the MD in mitigating microglia-mediated neuroinflammation, including explanation regarding their bioavailability, metabolism and blood–brain barrier.

La Autoridad Europea de Seguridad Alimentaria (EFSA) es el principal organismo europeo encargado de la Seguridad Alimentaria. En 2011, elaboró una opinión científica sobre la formación de aminas biógenas en alimentos fermentados, en la que se pone de manifiesto la importancia del control de estos compuestos causantes de reacciones adversas en los consumidores. Así, es preciso disponer de mayor conocimiento científico en relación a la presencia de aminas biógenas en alimentos fermentados para realizar una mejor evaluación del riesgo, especialmente en la disponibilidad de métodos de análisis adecuados. Por el contrario, la EFSA también es la encargada de evaluar las Declaraciones de Propiedades Saludables de los alimentos. En este caso, para demostrar una declaración saludable hay que caracterizar el alimento, definir su actividad fisiológica, así como su disponibilidad y plausibilidad biológica. Por todo ello, para poder evaluar las funciones biológicas de los compuestos fenólicos es imprescindible conocer su biodisponibilidad. Además, es importante evaluar e identificar las transformaciones de estos compuestos a lo largo del tracto gastrointestinal, ya que se ha observado que las propiedades bioactivas de los metabolitos, aun siendo los principales implicados en los efectos beneficiosos, suelen ser diferentes a la de sus compuestos originales presentes en los alimentos. El objetivo de la presente Tesis Doctoral ha sido el desarrollo y validación de nuevos métodos analíticos basados en técnicas cromatográfícas para caracterizar y cuantificar compuestos con actividad biológica, como son las aminas biógenas, compuestos fenólicos y sus derivados, en productos vegetales, bebidas fermentadas y muestras biológicas obtenidas de estudios de biodisponibilidad. Estos nuevos métodos han sido aplicados a la resolución de problemas agrupados fundamentalmente en dos grandes bloques temáticos, Seguridad alimentaria y Calidad Alimentaria. La primera parte de la memoria de esta Tesis

La presente Tesis Doctoral se centra en el análisis de compuestos bioactivos en productos fermentados. Una primera parte ha tenido como objeto la determinación de los polifenoles no antocianos en un puré de fresa y en los fermentados alcohólicos y glucónicos obtenidos a partir de este sustrato. También se realizó el estudio de estabilidad de una bebida obtenida a partir de la fermentación glucónica. Dicha bebida fue diseñada como una alternativa viable para el aprovechamiento de los excedentes de producción de una fruta de marcado carácter perecedero y de elevada producción en España. La segunda parte se ha centrado en la capacidad de las levaduras vínicas como productoras de compuestos bioactivos a partir de determinados aminoácidos durante el proceso de fermentación alcohólica. A tal fin, se han identificado y determinado los polifenoles no antocianos mediante el empleo de diferentes técnicas y equipos, como HPLC-DAD, UPLC-QTRAP (QqQLIT) y UPLCQExactive. En todos los análisis de espectrometría de masas se utilizó el método MRM (Multiple Reaction Monitoring). Esta tarea identificativa tuvo como objeto el análisis del puré de fresa utilizado posteriormente como sustrato para la elaboración de dos fermentados diferentes (glucónico y alcohólico). El resultado de esta actividad nos permitió identificar un total de 106 compuestos fenólicos, 32 en el puré de fresa, 43 en los fermentados glucónicos, 66 en las fermentaciones alcohólicas y 64 en la bebida. Todo ello facilitó la identificación de 25 compuestos que, a la fecha de las publicaciones, no habían sido aún descritos en la fresa o productos derivados (12 flavonoles, 6 ácidos hidroxicinámicos, 2 taninos, 2 chalconas, 1 ácido hidroxibenzoico, 1 estilbeno y el ácido homovanillico). La valoración del impacto del proceso de fabricación industrial del puré de fresa sobre el contenido de los polifenoles no antocianos, permitió comprobar que este proceso es respetuoso con la composición de la fruta, preservando su capacidad, This Doctoral Thesis focuses on the analysis of bioactive compounds in fermented foods. The objective of the first part is to determine the non-anthocyanin polyphenols in the strawberry purée and in the gluconic and alcoholic fermented products obtained from this substrate. We also studied the stability of a drink elaborated from gluconic fermentation that was used to produce a drink designed as a viable alternative to take advantage of the surplus of the production of this perishable fruit so produced in Spain. The second part focuses in the ability of the winemaking yeast to produce the bioactive compounds in the alcoholic fermentation process. In order to do so, we have identified the composition of the non-anthocyanin polyphenols by using different techniques and instruments such as the HPLC-DAD, the UHPLCQTRAP (QqQLIT) and the UHPLC-QExactive. The method MRM (Multiple Reaction Monitoring) was used for all spectrometric mass analysis. The purpose was to analyse the strawberry purée used later as a substrate to elaborate those two different fermented products (gluconic and alcoholic). The outcome of this activity allowed us to identified 106 compounds, of which 32 were in the strawberry purée, 43 in the gluconic fermented product, 66 in the alcoholic fermented product and 64 in the drink. All of this enabled us to identify 25 compounds which had never been referred to before, to this date, in the strawberry purée or its derivatives (12 flavonols, 6 hydroxycinnamic acids, 2 tannins, 2 chalcones, 2 hydroxybenzoic acid, 1 stilbene and the homovanillic acid). The study of the impact of the process of industrial production of the strawberry purée in the content of the non-anthocyanin compound, allowed us to prove that this process preserves most of the composition of the fruit and its antioxidant properties. Another subject treated in the Thesis focused on the use of this strawberry purée as a substrate for the fermentation processes. The analysis of the processes sho

En los últimos años, la melatonina ha sido identificada en diversas matrices alimentarias, principalmente de origen vegetal como frutas (uvas, fresas) y verduras (tomates, pimientos). Sin embargo, ha generado un notable interés el hecho de detectar melatonina en alimentos fermentados como el vino o la cerveza. Todo parece indicar que la presencia de esta molécula en alimentos fermentados está directamente relacionada con el metabolismo de las levaduras, especialmente del género Saccharomyces cerevisiae, durante los procesos fermentativos. La melatonina es bien conocida en mamíferos como neurohormona sintetizada por la glándula pineal y su función principal es la de regular los ciclos circadianos de sueño/vigilia dependiendo de las condiciones de luz y oscuridad. Además, también ha mostrado otras propiedades bioactivas como molécula antioxidante, siendo una excelente captadora de especies reactivas de oxígeno. Por otro lado, ha sido también descrita en plantas y en microorganismos, aunque en este caso sus funciones no están completamente dilucidadas. La melatonina se forma a partir del aminóacido triptófano, que pertenece al grupo de aminoácidos aromáticos junto a la tirosina y la fenilalanina. Su proceso de formación en mamíferos contempla como intermediarios moléculas como el 5-hidroxitriptófano, la serotonina o la N-acetilserotonina. En plantas, la triptamina también puede formar parte de la ruta de formación de la melatonina en lugar del 5-hidroxitriptófano. Sin embargo, en microorganismos no está claramente definida su ruta metabólica de síntesis. En el caso de las levaduras, el metabolismo del triptófano produce asimismo el ácido indolacético y el triptofol, derivados de la ruta de Ehrlich. Las técnicas analíticas para cuantificar melatonina en fluidos biológicos y alimentos abarcan desde el radioinmunoensayo, en el que destaca la técnica ELISA, hasta las técnicas de cromatografía líquida de alta resolución con detectores de masas de última generación. El princ, In recent years, melatonin has been identified in different foods, mainly from plant origin such us fruits (grapes, strawberries) and vegetables (tomatoes and peppers). However, the identification of melatonin in fermented foods as beer and wine has attracted an increasing interest in research. Literature data indicate that the presence of this molecule in fermented food is due to the yeast metabolism, especially from Saccharomyces cerevisiae, during fermentative process. Melatonin is a well-known neurohormone synthesized by the pineal gland of mammals. Its main function is related with circadian rhythms depending of light/darkness conditions. Besides, it has reported additional bioactive properties such as antioxidant, being an effective oxygen scavenger. On the other hand, it has also been described in plants and microorganism, although its role in both organism in not well elucidated. Melatonin is formed from the amino acid tryptophan that belongs to the aromatic amino acids groups. Mammal’s metabolism of tryptophan encompasses different secondary metabolites such as 5-hydroxytryptophan, serotonin and N-acetylserotonin. In plants, tryptamine can be included on the melatonin pathway instead of 5-hydroxytryptophan. However, the metabolic pathway that produce melatonin in microorganisms is not completely defined. Nevertheless, amino acid catabolism by yeast known as the Ehrlich pathway mainly produces indole acetic acid and tryptophol. Analytical techniques to quantify melatonin in biological fluids and foodstuff is quite diverse, covering from radioimmunoassay, being ELISA the most representative, to high resolution liquid chromatographic techniques coupled to a high resolution mass spectrometer. The main challenge of these techniques is to improve the detection and quantification limits of these compounds. ELISA technique is well performed when biological fluids are analysed. Nevertheless, its application to foodstuffs has been less repeatable, maybe due to the pr

Hydroxytyrosol (HT) is a phenolic compound of recognized bioactivity that has been described in wines but little is known about its origin. This work demonstrates that yeast involved in wine making, i.e. Saccharomyces cerevisiae strains and the non-Saccharomyces Torulaspora delbrueckii, can synthesise HT, as this compound was identified in the intracellular media of three strains by means of a developed and validated UHPLC–HRMS method with LOQ and LOD of 0.108 and 0.035 ng mL−1 respectively. Controlled fermentations were performed with different varieties of grapes (Corredera, Moscatel, Chardonnay, Palomino fino, Sauvignon Blanc, Vijiriega, and Tempranillo) and synthetic must. The Saccharomyces cerevisiae strain QA23 was the most efficient producer of HT from tested yeasts. On the other hand, the grape variety influences HT wine concentrations. Furthermore, the maximum concentration of HT is reached between the fourth and sixth day of fermentation. This work reveals that yeasts have a great potential for the production of HT.

El sobrepeso y la obesidad, actualmente consideradas como pandemia a nivel mundial, han ido incrementando con el paso de las décadas. Ambas enfermedades presentan altas tasas de morbilidad y mortalidad, ya que se les asocian complicaciones tales como dislipemia, diabetes e hipertensión entre otras. La falta de buenos hábitos alimentarios y el sedentarismo son los principales motivos de este peligroso incremento. En este contexto las aplicaciones móviles y web se ofrecen como un medio que permite la evaluación de la dieta por parte del propio usuario y como herramienta para dar y recibir nociones sobre alimentación. El objetivo de este trabajo ha sido evaluar la eficacia del uso de una de estas apps, Balanceat, para mejorar la dieta de un grupo de personas, consiguiendo que sea más equilibrada. Dado que el farmacéutico ejerce consejo dietético, se han seleccionado a estudiantes de la asignatura optativa de Nutrición, Dietética y Dietoterapia del Grado en Farmacia impartida en la Facultad de Farmacia de la Universidad de Sevilla para que además de evaluar la herramienta, esta intervención sirviera para reforzar esta competencia mediante el uso de las nuevas tecnologías. Por ello, en primer lugar, los estudiantes debían evaluar su dieta y después, realizar consejo dietético a una persona con sobrepeso. En este trabajo, se presentan los resultados de estos dos grupos evaluándose la dieta antes y después de uso de la plataforma. Ambos grupos mostraron una mejora satisfactoria de sus hábitos alimentarios, tanto calórica como en la distribución de macronutrientes y número de comidas realizadas al día. En base a estos resultados, se puede afirmar que utilizando la app Balanceat, los individuos han mejorado sus hábitos hacia una alimentación más saludable.

Excessive concentrations of vascular endothelial growth factor (VEGF) trigger angiogenesis, which causes complications such as the destabilization of atherosclerotic plaques and increased growth of tumors. This work focuses on the determination of the inhibitory activity of melatonin and other indolic related compounds on VEGF-induced VEGF receptor-2 (VEGFR-2) activation and an approximation to the molecular mechanism underlying the inhibition. Quantification of phosphorylated VEGFR-2 was measured by ELISA. Migration wound-healing assay was used to determine cell migration of human umbilical vein endothelial cells (HUVECs). This is the first time that melatonin, 3-indolacetic acid, 5-hydroxytryptophol, and serotonin are proved to significantly inhibit VEGF-induced VEGFR-2 activation in human umbilical vein endothelial cells and subsequent angiogenesis. 3-Indolacetic acid showed the highest inhibitory effect (IC50 value of 0.9704 mM), followed by 5-hydroxytryptophol (35% of inhibition at 0.1 mM), melatonin (30% of inhibition at 1 mM), and serotonin (24% of inhibition at 1 mM). An approximation to the molecular mechanism of the inhibition has been proposed, suggesting that indolic compounds might interact with the cell surface components of the endothelial membrane in a way that prevents VEGF from activating the receptor. Additionally, wound-healing assay revealed that exposure of HUVECs to melatonin and 3-indolacetic acid in the presence of VEGF significantly inhibited cell migration by 87% and 99%, respectively, after 24 h. These data demonstrate that melatonin, 3-indolacetic acid, 5-hydroxytryptophol, and serotonin would be good molecules for future exploitation as anti-VEGF signaling agents.

La fresa es una excelente fuente de compuestos bioactivos, sin embargo, es un fruto muy perecedero. La elaboración de productos fermentados supone una valiosa estrategia para disminuir las pérdidas económicas por excedentes de fruta y aumentar su vida útil. Por otro lado, resulta de gran interés el desarrollo de procesos biotecnológicos que preserven los compuestos bioactivos presentes en el fruto de partida. En esta Tesis Doctoral se ha evaluado la composición antociánica, color y actividad antioxidante de productos fermentados elaborados a partir de puré de fresa, concretamente, fermentados alcohólicos, acéticos y glucónicos, así como la estabilidad de una bebida obtenida a partir de estos productos. Asimismo, se ha estudiado el efecto neuroprotector de estos fermentados frente a la toxicidad del péptido β-amiloide (Aβ) y a la producción de Especies Reactivas de Oxígeno (ROS). Por otro lado, se ha demostrado la bioactividad del ácido protocatéquico (uno de los principales metabolitos de antocianos) frente a la agregación y toxicidad del péptido Aβ y de la proteína α-sinucleína (αS) in vitro. Además, se ha analizado el efecto de otros compuestos bioactivos (melatonina y otros compuestos indólicos) presentes en fresa y otros productos fermentados frente al proceso de fibrilación y neurotoxicidad del péptido Aβ. Los resultados obtenidos han permitido la identificación de nuevos compuestos antociánicos en fermentados alcohólicos y acéticos. Al mismo tiempo, se ha comprobado cómo el proceso que mejor preserva los antocianos es la fermentación glucónica, que se ha seleccionado como base de la bebida elaborada. Además, los fermentados alcohólicos y glucónicos protegen a las células neuronales de la toxicidad del péptido Aβ. Asimismo, el ácido protocatéquico posee un efecto inhibidor de la formación de fibras de Aβ y αS y efectos neuroprotectores demostrado por diversas técnicas in vitro. Por último, el estudio de los compuestos indólicos frente al proceso de agregación y, The strawberry is an excellent source of bioactive compounds, however, it is a very perishable fruit. The elaboration of fermented products supposes a valuable strategy to prevent the economic losses due to strawberry surplus and to increase the shelf life. On the other hand, the use of biotechnological processes that preserve the bioactive compounds is a relevant aspect to take into consideration. In this PhD Thesis, anthocyanin composition, colour and antioxidant activity of different fermented products made from strawberry purée by alcoholic, acetic and gluconic fermentations have been evaluated. Additionally, the stability of a beverage obtained from these products has been studied. Moreover, the neuroprotective effect of these products against amyloid-β (Aβ) toxicity and Reactive Oxygen Species (ROS) production has been investigated. In addition, the bioactivity of protocatechuic acid (one of the most important anthocyanin metabolite) against the aggregation and toxicity of Aβ peptide and α-synuclein (αS) protein in vitro has been demonstrated. Furthermore, it has been confirmed that other bioactive compounds (melatonin and indolic related compounds) described in strawberry and in other fermented products present an effect on the inhibition of the Aβ aggregation and the Aβ-induced toxicity. The obtained results have led to the identification of new anthocyanin compounds in alcoholic and acetic fermented products. Moreover, it has been proven that gluconic fermentation is the process, from those studied in this Thesis, that better preserves the anthocyanin profile. Thus, it was the one selected as substrate to elaborate the beverage. Furthermore, alcoholic and gluconic fermented extracts protect neuronal cells against Aβ toxicity. Protocatechuic acid possesses an inhibitory effect of the Aβ and αS fibril formation and neuroprotective properties demonstrated by several in vitro techniques. Finally, serotonin has been proved to be a potent inhibitor of the fibril

Metabolites resulting from nitrogen metabolism in yeast are currently found in some fermented beverages such as wine and beer. Their study has recently attracted the attention of researchers. Some metabolites derived from aromatic amino acids are bioactive compounds that can behave as hormones or even mimic their role in humans and may also act as regulators in yeast. Although the metabolic pathways for their formation are well known, the physiological significance is still far from being understood. The understanding of this relevance will be a key element in managing the production of these compounds under controlled conditions, to offer fermented food with specific enrichment in these compounds or even to use the yeast as nutritional complements

Polyphenolic compounds, widely distributed in plant kingdom, have been exhaustively studied for their bioactive properties specially antioxidant activity. However, they are extensively metabolized by human organism and the resulting metabolites are largely responsible for their effects. Furthermore, they may interact with the endogenous antioxidant network being this possibility scarcely studied. Plasma antioxidant network encompasses antioxidant enzymes and other substances such as uric acid. In addition, ascorbic acid is the major compound representing water soluble compartment both in foods and human body. The interaction of this vitamin with phenolic compound is largely unexplored. This work aims to study if there is a synergic effect between phenolic metabolites and main antioxidants (uric and ascorbic acid). For this purpose, the antioxidant activity was evaluated in terms of ORAC (oxygen radical absorbance capac ity) and FRAP (ferricreducing antioxidant power) as these tests involved either HAT (Hydrogen Atom Transfer) or SET (Single Electron Transfer) mechanisms. Additionally, a kinetic studied was developed to test if the rate constant presented a synergic effect. Protocatechuic acid, 3, 4 -dihydroxyphenylacetic acid, 3,4 -dihydroxyphenylpropionic acid and 3 -hydroxyphenylacetic acid were selected as they were metabolites of polyphenol compounds such as anthocyanins, quercetin, neohesperidin, chlorogenic acid and hesperetin present in wines, orange and strawberries. A synergic effect was proved for the combination of ascorbic acid with 3,4 -dihydroxyphenylacetic acid, both in terms of antioxidant activity and potent increase of velocity of the antioxidant reaction that took place.

A portion of the odourless fraction of fruits bound to sugars releases aromatic substances that represent an important source of aromatic potential. During the processing of fruits, these compounds may be affected. Thus, in this work, for the first time, glycosidic aroma precursors were studied over the course of the industrial production process of commercial strawberry puree. Free volatile compounds were also studied. The results indicated that the amounts of free and bound aromatic compounds decreased, particularly in the free fraction, by more than 50% of the total amounts. The pasteurisation stage led to the greatest loss of the precursors of key strawberry odorants. However, the seed removal step offset these losses during the processing of glycosidically bound aroma compounds. The free volatile compounds that were most strongly affected were the higher alcohols and ethyl esters. This study suggests that the amounts of glycosidic aroma precursors in the raw material significantly affect the aromatic potentials of commercial purees.

La presente invención tiene por objeto la aromatización con extractos naturales de nectarina, vainilla y lima de vinos con bajo contenido en alcohol, con el fin de obtener productos aceptables sensorialmente por los consumidores.